LOCUS       sequence-415695-        5500 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expression of Cav2.1 N-terminus (amino acids 1-100) with a 
            C-terminal CAAX (last 10aa of H.Ras) motif and a HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5500)
  AUTHORS   Page KM, Heblich F, Margas W, Pratt WS, Nieto-Rostro M, Chaggar K, 
            Sandhu K, Davies A, Dolphin AC
  TITLE     N terminus is key to the dominant negative suppression of Ca(V)2 
            calcium channels: implications for episodic ataxia type 2.
  JOURNAL   J Biol Chem. 2010 Jan 8;285(2):835-44. doi: 10.1074/jbc.M109.065045.
            Epub 2009 Nov 10.
  PUBMED    19903821
REFERENCE   2  (bases 1 to 5500)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M109.065045"; journalName: "J Biol Chem"; date: 
            "2010-01-8- 8"; volume: "285"; issue: "2"; pages: "835-44"
COMMENT     Sequence Label: An HA CAAX pMT2
FEATURES             Location/Qualifiers
     source          1..5500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..36)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(696..1487,1488..1556))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1319..1338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1557..1661)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1729..1747
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1785..1807)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1907..1926
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2120..2142
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2134..2151
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2135..2151
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2161..2490)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2175..2310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2475..2495
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          2959..3188
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3171..3190
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3548..3574
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3709..4272
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    4459..4593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4496..4515)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4550..4569
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5266..5283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5437..5500,1..525))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
       61 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      121 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      181 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      241 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      301 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      361 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      421 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      481 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      541 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      601 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      661 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      721 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      781 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      841 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      901 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      961 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1021 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1081 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1141 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1201 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1261 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1321 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1381 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1441 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1501 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1561 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1621 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1681 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     1741 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     1801 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     1861 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     1921 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     1981 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2041 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2101 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
     2161 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     2221 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     2281 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     2341 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     2401 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     2461 tggggacttt ccacacccta actgacacac attccacagg atccggtcgc gcgaatttcg
     2521 agcggtgttc cgcggtcctc ctcgtataga aactcggacc actctgagac gaaggctcgc
     2581 gtccaggcca gcacgaagga ggctaagtgg gaggggtagc ggtcgttgtc cactaggggg
     2641 tccactcgct ccagggtgtg aagacacatg tcgccctctt cggcatcaag gaaggtgatt
     2701 ggtttatagg tgtaggccac gtgaccgggt gttcctgaag gggggctata aaagggggtg
     2761 ggggcgcgtt cgtcctcact ctcttccgca tcgctgtctg cgagggccag ctgttgggct
     2821 cgcggttgag gacaaactct tcgcggtctt tccagtactc ttggatcgga aacccgtcgg
     2881 cctccgaacg gtactccgcc accgagggac ctgagcgagt ccgcatcgac cggatcggaa
     2941 aacctctcga ctgttggggt gagtactccc tctcaaaagc gggcatgact tctgcgctaa
     3001 gattgtcagt ttccaaaaac gaggaggatt tgatattcac ctggcccgcg gtgatgcctt
     3061 tgagggtggc cgcgtccatc tggtcagaaa agacaatctt tttgttgtca agcttgaggt
     3121 gtggcaggct tgagatctgg ccatacactt gagtgacaat gacatccact ttgcctttct
     3181 ctccacaggt gtccactccc aggtccaact gcaggcgagc ctgaattgcg gccgcactag
     3241 tatggcccgc tttggagacg agatgccggg ccgctacggc gcaggcggag gaggctcagg
     3301 gccggccgcc ggggtggtcg tgggcgccgc gggcggccga ggagccgggg gcagccggca
     3361 gggcgggcag cccggagcgc agaggatgta caagcagtcg atggcgcaga gagcgcggac
     3421 catggccctc tacaacccca tccctgtccg ccagaactgc ctcacggtca accgctccct
     3481 gttcctcttc agtgaagaca acgtggtgag aaaatacgcc aaaaagatca cggaatggcc
     3541 tcgagtctac ccttacgacg tccctgacta cgccgttcga ggctgcatga gctgcaagtg
     3601 tgtgctctcc tgaattccgg ggggggacag ctcagggctg cgatttcgcg ccaaacttga
     3661 cggcaatcct agcgtgaagg ctggtaggat tttatccccg ctgccatcat ggttcgacca
     3721 ttgaactgca tcgtcgccgt gtcccaaaat atggggattg gcaagaacgg agacctaccc
     3781 tggcctccgc tcaggaacga gttcaagtac ttccaaagaa tgaccacaac ctcttcagtg
     3841 gaaggtaaac agaatctggt gattatgggt aggaaaacct ggttctccat tcctgagaag
     3901 aatcgacctt taaaggacag aattaatata gttctcagta gagaactcaa agaaccacca
     3961 cgaggagctc attttcttgc caaaagtttg gatgatgcct taagacttat tgaacaaccg
     4021 gaattggcaa gtaaagtaga catggtttgg atagtcggag gcagttctgt ttaccaggaa
     4081 gccatgaatc aaccaggcca cctcagactc tttgtgacaa ggatcatgca ggaatttgaa
     4141 agtgacacgt ttttcccaga aattgatttg gggaaatata aacttctccc agaataccca
     4201 ggcgtcctct ctgaggtcca ggaggaaaaa ggcatcaagt ataagtttga agtctacgag
     4261 aagaaagact aacaggaaga tgctttcaag ttctctgctc ccctcctaaa gctatgcatt
     4321 tttataagac catgggactt ttgctggctt tagatcataa tcagccatac cacatttgta
     4381 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4441 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4501 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4561 aaactcatca atgtatctta tcatgtctgg atccccggcc aacggtctgg tgacccggct
     4621 gcgagagctc ggtgtacctg agacgcgagt aagcccttga gtcaaagacg tagtcgttgc
     4681 aagtccgcac caggtactga tatcccacca aaaagtgcgg cggcggctgg cggtagaggg
     4741 gccagcgtag ggtggccggg gctccggggg cgaggtcttc caacataagg cgatgatatc
     4801 cgtagatgta cctggacatc caggtgatgc cggcggcggt ggtggaggcg cgcggaaagt
     4861 cgcggacgcg gttccagatg ttgcgcagcg gcaaaaagtg ctccatggtc gggacgctct
     4921 ggccggtgag gcgtgcgcag tcgttgacgc tctagaccgt gcaaaaggag agcctgtaag
     4981 cgggcactct tccgtggtct ggtggataaa ttcgcaaggg tatcatggcg gacgaccggg
     5041 gttcgaaccc cggatccggc cgtccgccgt gatccatgcg gttaccgccc gcgtgtcgaa
     5101 cccaggtgtg cgacgtcaga caacggggga gcgctccttt tggcttcctt ccaggcgcgg
     5161 cggctgctgc gctagctttt ttggcgagct cgaattaatt ctgcattaat gaatcggcca
     5221 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5281 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5341 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5401 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
//
